DAXX 通过激活 ERK 信号通路促进卵巢癌腹水细胞的增殖和迁移。
DAXX promotes ovarian cancer ascites cell proliferation and migration by activating the ERK signaling pathway.
机构信息
College of Medicine, Jiaxing University, Jiaxing, 314001, China.
出版信息
J Ovarian Res. 2018 Oct 18;11(1):90. doi: 10.1186/s13048-018-0462-4.
BACKGROUND
The death-domain-associated protein (DAXX) was originally identified as a protein that binds to the transmembrane death receptor FAS and enhances both FAS-induced and transforming growth factor-β-dependent apoptosis. In a previous study, we found that nude mice injected with DAXX-overexpressing cells (ES-2-DAXX) accumulated large concentrations of first-generation ascites cells (I ascites cells). The role of DAXX in the development of ascites is unknown. The aim of this study was to analyze the effect of DAXX on proliferation and migration of ascites cells in ovarian cancer in vitro and in vivo.
METHODS
Nude mice were housed in cages with a 14:10 h light:dark cycle; water and food were provided ad libitum. ES-2-DAXX cells (1×106) were injected intraperitoneally into athymic nude mice (8-week-old female mice). After 4 weeks, I ascites cells were collected. The I ascites cells were injected intraperitoneally into athymic nude mice (8-week-old female mice). After 4 weeks, II ascites cells were collected and cultured. Ascites cell survival, migration, and colony formation were measured using colony formation and cell growth assays. Immunofluorescent staining revealed the co-localization of DAXX and promyelocytic leukemia protein (PML) in ascites cell nuclei. Western blotting and immunohistochemistry showed that extracellular signal-related kinase (p-ERK) 1/2 and CEBP-β were highly expressed in tumor tissues formed by II ascites cells. Through immunoprecipitation, we also found that DAXX can interact with CEBP-β.
RESULTS
DAXX enhanced ascites cell survival, migration, and colony formation. DAXX and PML nuclear foci dramatically increased in a passage-dependent manner in ascites cells, DAXX promoted the tumor growth of ascites cells in vivo, increased ascites cell proliferation in vivo, and enhanced ascites cell survival and migration by activating the ERK signalling pathway and integrating with CEBP-β.
CONCLUSIONS
DAXX can interact with CEBP-β. DAXX can induce ovarian cancer ascites formation by activating the ERK signal pathway and binding to CEBP-β.
背景
死亡结构域相关蛋白（DAXX）最初被鉴定为一种与跨膜死亡受体 Fas 结合的蛋白，可增强 Fas 诱导的和转化生长因子-β依赖性凋亡。在之前的研究中，我们发现过表达 DAXX 的细胞（ES-2-DAXX）注射入裸鼠后，裸鼠积累了大量第一代腹水细胞（I 型腹水细胞）。DAXX 在腹水形成中的作用尚不清楚。本研究旨在分析 DAXX 对卵巢癌腹水细胞在体外和体内增殖和迁移的影响。
方法
裸鼠饲养在 14:10 h 光：暗循环的笼中；提供自由饮水和食物。将 ES-2-DAXX 细胞（1×106）注射入 8 周龄雌性裸鼠腹腔内。4 周后，收集 I 型腹水细胞。将 I 型腹水细胞注射入 8 周龄雌性裸鼠腹腔内。4 周后，收集 II 型腹水细胞并培养。通过集落形成和细胞生长测定法测量腹水细胞的存活、迁移和集落形成。免疫荧光染色显示 DAXX 和早幼粒细胞白血病蛋白（PML）在腹水细胞核中的共定位。Western blot 和免疫组化显示，II 型腹水细胞形成的肿瘤组织中细胞外信号调节激酶（p-ERK）1/2 和 CCAAT 增强子结合蛋白-β（CEBP-β）高度表达。通过免疫沉淀，我们还发现 DAXX 可以与 CEBP-β 相互作用。
结果
DAXX 增强了腹水细胞的存活、迁移和集落形成。DAXX 和 PML 核焦点在腹水细胞中呈依赖性增加，DAXX 促进了腹水细胞在体内的肿瘤生长，增强了体内腹水细胞的增殖，并通过激活 ERK 信号通路和与 CEBP-β 整合，增强了腹水细胞的存活和迁移。
结论
DAXX 可以与 CEBP-β 相互作用。DAXX 通过激活 ERK 信号通路并与 CEBP-β 结合，诱导卵巢癌腹水形成。
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